April 19, 2024

Alternative solutions to separate AAV full and empty capsids using anion exchange

Adeno-associated virus (AAV) is the main vector for gene therapy and there is need for scalable, cost-efficient, and robust chromatography-based purification. Key for a successful process are high overall yields of full capsids with effective empty capsid reduction and efficient impurity removal.

Here we present high-performance anion exchange (AEX) chromatography separation for several AAV serotypes using step-elution mode. Optimized protocols and conditions for both resin and membrane formats are described. Full and empty capsid peaks were identified using the UV260:280 ratio from chromatograms. And separation results were confirmed by PCR, ELISA, mass photometry, and analytical ultracentrifugation. We show that these AEX approaches are reproducible and can be scaled up.

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Tags: membranes, resins, downstream, chromatography, purification, viral vector, AAV